RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana

Abstract Background The Arabidopsis RUS (ROOT UV-B SENSITIVE) gene family contains six members, each of which encodes a protein containing a DUF647 (domain of unknown function 647) that is commonly found in eukaryotes. Previous studies have demonstrated that RUS1 and RUS2 play critical roles in earl...

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Main Authors: Nathaniel Perry, Colin D. Leasure, Hongyun Tong, Elias M. Duarte, Zheng-Hui He
Format: Article
Language:English
Published: BMC 2021-05-01
Series:BMC Plant Biology
Subjects:
Online Access:https://doi.org/10.1186/s12870-021-03011-8
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spelling doaj-1fa187e04957423ba17ac8211e53ab752021-05-30T11:19:33ZengBMCBMC Plant Biology1471-22292021-05-0121111410.1186/s12870-021-03011-8RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thalianaNathaniel Perry0Colin D. Leasure1Hongyun Tong2Elias M. Duarte3Zheng-Hui He4Department of Biology, San Francisco State UniversityDepartment of Biology, San Francisco State UniversityDepartment of Biology, San Francisco State UniversityDepartment of Biology, San Francisco State UniversityDepartment of Biology, San Francisco State UniversityAbstract Background The Arabidopsis RUS (ROOT UV-B SENSITIVE) gene family contains six members, each of which encodes a protein containing a DUF647 (domain of unknown function 647) that is commonly found in eukaryotes. Previous studies have demonstrated that RUS1 and RUS2 play critical roles in early seedling development. All six RUS genes are expressed throughout the plant, but little is known about the functional roles of RUS3, RUS4, RUS5 and RUS6. Results We used a reverse-genetic approach to identify knockout mutants for RUS3, RUS4, RUS5 and RUS6. Each mutant was confirmed by direct DNA sequencing and genetic segregation analysis. No visible phenotypic differences were observed in rus3, rus4, or rus5 knockout mutants under standard growth conditions, but rus6 knockout mutants displayed a strong embryo-lethal phenotype. Two independent knockout lines for RUS6 were characterized. The rus6 mutations could only be maintained through a heterozygote, because rus6 homozygous mutants did not survive. Closer examinations of homozygous rus6 embryos from rus6/ + parent plants revealed that RUS6 is required for early embryo development. Loss of RUS6 resulted in embryo lethality, specifically at the mid-globular stage. The embryo-lethality phenotype was complemented by a RUS6::RUS6-GFP transgene, and GFP signal was detected throughout the embryo. Histological analyses with the β-glucuronidase reporter gene driven by the RUS6 promoter showed tissue- and development-specific expression of RUS6, which was highest in floral tissues. Conclusion Our data revealed that RUS6 is essential for early embryo development in Arabidopsis, and that the RUS gene family functions in multiple stages of plant development.https://doi.org/10.1186/s12870-021-03011-8RUS Gene FamilyDUF647Embryo DevelopmentComplementationArabidopsis
collection DOAJ
language English
format Article
sources DOAJ
author Nathaniel Perry
Colin D. Leasure
Hongyun Tong
Elias M. Duarte
Zheng-Hui He
spellingShingle Nathaniel Perry
Colin D. Leasure
Hongyun Tong
Elias M. Duarte
Zheng-Hui He
RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
BMC Plant Biology
RUS Gene Family
DUF647
Embryo Development
Complementation
Arabidopsis
author_facet Nathaniel Perry
Colin D. Leasure
Hongyun Tong
Elias M. Duarte
Zheng-Hui He
author_sort Nathaniel Perry
title RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
title_short RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
title_full RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
title_fullStr RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
title_full_unstemmed RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana
title_sort rus6, a duf647-containing protein, is essential for early embryonic development in arabidopsis thaliana
publisher BMC
series BMC Plant Biology
issn 1471-2229
publishDate 2021-05-01
description Abstract Background The Arabidopsis RUS (ROOT UV-B SENSITIVE) gene family contains six members, each of which encodes a protein containing a DUF647 (domain of unknown function 647) that is commonly found in eukaryotes. Previous studies have demonstrated that RUS1 and RUS2 play critical roles in early seedling development. All six RUS genes are expressed throughout the plant, but little is known about the functional roles of RUS3, RUS4, RUS5 and RUS6. Results We used a reverse-genetic approach to identify knockout mutants for RUS3, RUS4, RUS5 and RUS6. Each mutant was confirmed by direct DNA sequencing and genetic segregation analysis. No visible phenotypic differences were observed in rus3, rus4, or rus5 knockout mutants under standard growth conditions, but rus6 knockout mutants displayed a strong embryo-lethal phenotype. Two independent knockout lines for RUS6 were characterized. The rus6 mutations could only be maintained through a heterozygote, because rus6 homozygous mutants did not survive. Closer examinations of homozygous rus6 embryos from rus6/ + parent plants revealed that RUS6 is required for early embryo development. Loss of RUS6 resulted in embryo lethality, specifically at the mid-globular stage. The embryo-lethality phenotype was complemented by a RUS6::RUS6-GFP transgene, and GFP signal was detected throughout the embryo. Histological analyses with the β-glucuronidase reporter gene driven by the RUS6 promoter showed tissue- and development-specific expression of RUS6, which was highest in floral tissues. Conclusion Our data revealed that RUS6 is essential for early embryo development in Arabidopsis, and that the RUS gene family functions in multiple stages of plant development.
topic RUS Gene Family
DUF647
Embryo Development
Complementation
Arabidopsis
url https://doi.org/10.1186/s12870-021-03011-8
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